Western: wrong 2. Ab- R MY RESULTS USELESS?! - (Oct/10/2006 )

Hello everybody! I have a BIIIG problem. I used a wrong second antibody (rabbit instead of mouse) and -of yourse- faced problems while developing... still...I got some bands. Which even fit my predictions. And they have the right size. So: are these results reliable? Is a cross-reaction possible? Or should I just forget about it? And start all over again?

-Jou-

yes cross reaction is possible, however, why don't you incubate your membrane with the right antibody now? you would get nice bands.

-Missele-

Do I have to incubate with the first Antibody again prior to incubating with the right secondary Ab? or follow a certain washing procedure?Thank you!

-Jou-

QUOTE (Jou @ Oct 10 2006, 04:06 PM)

Do I have to incubate with the first Antibody again prior to incubating with the right secondary Ab? or follow a certain washing procedure?Thank you!

just wash the substrate, and incubate with the second antibody. the primary antibody is still bound, don't worry (unless all the blot strippers would have less problems !)